STUDIES OF THE DETERMINATION OF THE BACTERIUM COLI TYPE I CONTENT OF FARM WATER SUPPLIES

SUMMARY: A – comparison of the suitability of brilliant green bile broth and MacConkey's broth at 44° for the detection of Bacterium coli type I in farm water supplies, showed that 83.1% of the samples had no difference in the number of positive tubes at 44°, and only 5 samples (1.7%) had a significantly higher number of positive tubes in MacConkey's broth.
Of 707 strains of coli-aerogenes bacteria isolated from 44° positive tubes of both media, 94.5% were Bact. coli type I. Strains of Bact. coli type II and Bact. aerogenes type I which were 44° positive constituted 3.7% and 0.4% respectively, all of which were indole negative at 44°. In addition there were 10 strains (1.4%) of 44° positive Intermediate type II, 9 of which were indole positive at 44°.
An appreciable number (6.6%) of Bact. coli type I strains failed to give a positive indole reaction in 24 hr at 44°.     

THE BACTERIOLOGY OF FARM WATER SUPPLIES: A STUDY OF THE COLONY COUNT IN 72 HOURS AT 22°

SUMMARY: The colony count at 22° of farm water supplies from springs and wells was mainly composed of biochemically inactive, non-pigmented, Gram-negative rods. Water from a stream polluted with farmyard sewage showed a similar dominance of Gram-negative rods, but orange or yellow pigmented colonies were more abundant. There were few 37° positive coli-aerogenes bacteria in either the farm water supplies or the sewage polluted stream, and Bact. coli type I was rare.
A high proportion of the bacteria from farm water supplies fermented milk in 3 days at 22°; a third developed acid, 15% proteolysis and 6.4% ropiness.
Contamination of pure spring water with surface soil from a heavily grazed pasture resulted in a hundredfold increase in colony count with aerobic sporing rods replacing Gram-negative rods as the dominant organisms, but coli-aerogenes bacteria were absent.     

THE INCIDENCE OF BACTERIUM COLI IN FARM WATER SUPPLIES

SUMMARY: Farm water supplies giving 37° positives in MacConkey's broth within 24 hr. had a much higher incidence of presumptive Bact. coli type I than those which gave 37° positives during the second day of incubation only; presumptive Bact. coli reactions were obtained with 85% of the 7,522 tubes positive in 24 hr. compared with 23% for the 7,593 tubes showing positive reactions during the second day at 37°.     

THE DISINFECTANT PROPERTIES OF QUATERNARY AMMONIUM COMPOUNDS AND SODIUM HYPOCHLORITE: A COMPARISON OF RESULTS USING DIFFERENT TEST METHODS

SUMMARY: On exposing a strain of Bacterium coli 28.D.10 in a surface film at atmospheric temperature to atmospheres of different moisture contents, it was found that for relative humidities between 100 and 66% the numbers of survivors decreased with decreasing humidity. There was also some evidence of a slight increase in survivors for a decrease in relative humidity from 43 to 0%.
The percentage of survivors of Bact. coli after exposure to quaternary ammonium disinfectants also decreased with relative humidity between 100 and 66% but no significant differences were found for changes in relative humidity below 66%. The numbers of survivors of a culture of Staphylococcus aureus were the same after storage at a relative humidity of 43% as at 100%; drying did not appear to affect the sensitivity of Staph. aureus to quaternary ammonium compounds. Tests of the effect of storage time in a saturated atmosphere gave results which were not entirely consistent, but where differences were observed, there was a lower percentage of survivors for freshly inoculated films than for films which had been stored for 3 hr, presumably because a fresh film was more easily removed to the disinfectant.
When either Bact. coli or Staph. aureus was exposed to a disinfectant, the percentage of survivors was higher when the organisms were in a surface film than when they were inoculated directly into the disinfectant. Agitation during exposure reduced the numbers of survivors from a surface film. Neither the glass nor the metal coming in contact with the disinfectants affected the level of survivors.
Under the conditions of testing, sodium hypochlorite was a more effective disinfectant than the quaternary ammonium compounds used.     

SOME OBSERVATIONS ON THE BACTERIAL CONTENT OF WATER IN WATERCRESS BEDS

SUMMARY: Samples of water flowing into watercress beds had higher bacterial contents during the summer months although this was less marked among samples from deep underground sources. Outlet samples showed higher contents than inlet samples and the bacterial content varied markedly with season, being highest in summer.
The presumptive coli-aerogenes contents at 30°, 37° and 44° also showed a similar seasonal variation among samples from the outlets but the marked inferiority of samples other than from bores was no longer apparent after the water had flowed through the beds. The dominant types were Bact. aerogenes type I, pectate liquefiers, Bact. coli type I, and Intermediate type I.
The importance of the pectate liquefiers was confirmed by plating on pectate gel and by their isolation from these plates and from MacConkey's broth.     

MARINE BACTERIA OF CARDIGAN BAY. I. BACTERIA OF AN OFF-SHORE AREA

SUMMARY: A study of marine bacteria from a fixed off-shore area in Cardigan Bay showed there was little significance in the variation between numbers from different levels within 8 fathoms deep. There was a suggestion of seasonal variation in surface water in colony counts at 15–18°. In general, the bacterial content was highest over the summer months, but showed little direct correlation with temperature, pH or light.
Strains were mainly obligate halophytes and small Gram-negative rods predominated (96%). Nearly 50% of strains isolated liquefied gelatin; otherwise, biochemical reactions were weak. The proportion of pigmented strains was low.
Alien bacteria were present in irregularly varying numbers in all samples and showed no seasonal variation. Coli-aerogenes strains were recorded in small numbers, at all depths and in all seasons, but Bact. coli type I only rarely occurred.     

A Bacteriological Survey of the Domestic Environment

The results of a survey of the bacterial flora in many sites in 21 homes are discussed. In all areas both wet and dry, coagulase negative, Gram positive cocci and Bacillus spp. were found. Wet areas such as kitchen sinks and drains contained large numbers of Escherichia coli and sometimes Klebsiella pneumoniae , Citrobacter and Enterobacter spp. In toilet areas, little evidence was found of contamination with organisms of faecal origin. Of 47 samples taken from teacloths and towels 22 were contaminated with Staphylococcus aureus although the actual numbers of this organism were low. Pseudomonas aeruginosa was isolated from several sites in one home only.     

Characterization of the coliform and enteric bacilli in the environment of calves with colibacillosis

In the first part of the present study the coliform and enteric bacilli in the environment of calves with colibacillosis were examined. The occurrence, number, and pathogenic properties of Escherichia coli in barnyard soils were obtained from six cattle ranches. The O and K serogroups of E. coli isolates obtained from the feces of calves with colibacillosis born at these cattle ranches were determined, and their serotypes were compared with the E. coli O and K serotypes found in soils. The results showed a reservoir of potentially pathogenic E. coli in barnyard soils contaminated with bovine feces. For the second part of this study, 6 healthy calves and 51 calves with colibacillosis were studied. The numbers of total aerobic heterotrophic bacteria, total streptococci, fecal streptococci, total coliforms, and fecal coliforms in the feces of calves were determined. In addition, coliform and enteric bacilli from the feces of both healthy and diseased calves were identified, and their indole, methyl red, Voges-Proskauer, citrate (IMViC) types were described. In parallel, the IMViC types of coliform and enteric bacilli isolated from barnyard soils previously contaminated with bovine feces were compared with those isolated from uncontaminated soils. All fecal specimens were also examined for the presence of rotavirus. No significant effect on the numbers of the bacterial types was found. The results suggest that the predominant IMViC types found in the feces of calves with colibacillosis originate from the soil. From this study it is apparent that the occurrence, number, and survival of E. coli in barnyard soils is related to ranch husbandry and sanitary practices.     

Characterization of the coliform and enteric bacilli in the environment of calves with colibacillosis.

In the first part of the present study the coliform and enteric bacilli in the environment of calves with colibacillosis were examined. The occurrence, number, and pathogenic properties of Escherichia coli in barnyard soils were obtained from six cattle ranches. The O and K serogroups of E. coli isolates obtained from the feces of calves with colibacillosis born at these cattle ranches were determined, and their serotypes were compared with the E. coli O and K serotypes found in soils. The results showed a reservoir of potentially pathogenic E. coli in barnyard soils contaminated with bovine feces. For the second part of this study, 6 healthy calves and 51 calves with colibacillosis were studied. The numbers of total aerobic heterotrophic bacteria, total streptococci, fecal streptococci, total coliforms, and fecal coliforms in the feces of calves were determined. In addition, coliform and enteric bacilli from the feces of both healthy and diseased calves were identified, and their indole, methyl red, Voges-Proskauer, citrate (IMViC) types were described. In parallel, the IMViC types of coliform and enteric bacilli isolated from barnyard soils previously contaminated with bovine feces were compared with those isolated from uncontaminated soils. All fecal specimens were also examined for the presence of rotavirus. No significant effect on the numbers of the bacterial types was found. The results suggest that the predominant IMViC types found in the feces of calves with colibacillosis originate from the soil. From this study it is apparent that the occurrence, number, and survival of E. coli in barnyard soils is related to ranch husbandry and sanitary practices.     

Earthworms as vectors of Escherichia coli O157:H7 in soil and vermicomposts

Survival and movement of Escherichia coli O157:H7 in both soil and vermicompost is of concern with regards to human health. Whilst it is accepted that E. coli O157:H7 can persist for considerable periods in soils, it is not expected to survive thermophilic composting processes. However, the natural behavior of earthworms is increasingly utilized for composting (vermicomposting), and the extent to which earthworms promote the survival and dispersal of the bacterium within such systems is unknown. The faecal material produced by earthworms provides a ready supply of labile organic substrates to surrounding microbes within soil and compost, thus promoting microbial activity. Earthworms can also cause significant movement of organisms through the channels they form. Survival and dispersal of E. coli O157:H7 were monitored in contaminated soil and farmyard manure subjected to earthworm digestion over 21 days. Our findings lead to the conclusion that anecic earthworms such as Lumbricus terrestris may significantly aid vertical movement of E. coli O157 in soil, whereas epigeic earthworms such as Dendrobaena veneta significantly aid lateral movement within compost. Although the presence of earthworms in soil and compost may aid proliferation of E. coli O157 in early stages of contamination, long-term persistence of the pathogen appears to be unaffected.     

THE SELECTIVE ACTION OF THALLOUS ACETATE FOR LACTOBACILLI

SUMMARY: The effect of thallous acetate has been examined on the growth of 23 strains of lactobacilli representing 13 different species. All strains grew satisfactorily in both broth and agar media containing 0.1% of thallous acetate. Of the 18 other organisms also examined the growth of the streptococci was unsuppressed, staphylococci, a micrococcus, B. licheniformis and Pr. mirabilis were partially inhibited and strains of Bact. coli, B. cereus, Chr. prodigiosum and Ps. fluorescens were almost completely inhibited by that concentration. By its use a valuable selective medium for isolating lactobacilli from natural habitats of mixed flora was obtained, other organisms, with the exception of streptococci, being almost completely eliminated.     

Composition of Methane-Oxidizing Bacterial Communities as a Function of Nutrient Loading in the Florida Everglades

Agricultural runoff of phosphorus (P) in the northern Florida Everglades has resulted in several ecosystem level changes, including shifts in the microbial ecology of carbon cycling, with significantly higher methane being produced in the nutrient-enriched soils. Little is, however, known of the structure and activities of methane-oxidizing bacteria (MOB) in these environments. To address this, 0 to 10?cm plant-associated soil cores were collected from nutrient-impacted (F1), transition (F4), and unimpacted (U3) areas, sectioned in 2-cm increments, and methane oxidation rates were measured. F1 soils consumed approximately two-fold higher methane than U3 soils; additionally, most probable numbers of methanotrophs were 4-log higher in F1 than U3 soils. Metabolically active MOB containing pmoA sequences were characterized by stable-isotope probing using 10?% (v/v) (13)CH(4). pmoA sequences, encoding the alpha subunit of methane monooxygenase and related to type I methanotrophs, were identified from both impacted and unimpacted soils. Additionally, impacted soils also harbored type II methanotrophs, which have been shown to exhibit preferences for high methane concentrations. Additionally, across all soils, novel pmoA-type sequences were also detected, indicating presence of MOB specific to the Everglades. Multivariate statistical analyses confirmed that eutrophic soils consisted of metabolically distinct MOB community that is likely driven by nutrient enrichment. This study enhances our understanding on the biological fate of methane being produced in productive wetland soils of the Florida Everglades and how nutrient-enrichment affects the composition of methanotroph bacterial communities.     

Incidence and relationship of group D streptococci with other indicator organisms in meats.

Raw and processed meats were analyzed for presumptive group D streptococci using KF streptococcus agar. Counts were compared with coliform, presumptive Escherichia coli, and Enterobacteriaceae counts but no meaningful relationships were observed. Results indicated that group D streptococci and E. coli type I were principally contaminants from the packing plant, rather than at retail level. The predominating group D streptococcus in both beef and pork cuts was Streptococcus faecalis, while in processed meat (bologna), the predominating group D streptococci were Streptococcus faecium var. durans and Streptococcus faecium. Streptococcus bovis was not detected among the isolates from any meat samples. Marked differences were noted in numbers of group D streptococci in processed meat from different manufacturers. The results did not support the use of group D streptococci as alternative indicator organisms for meats. However, the association of group D streptococci with packing plant contamination may prove to be of value.     

Deoxyribonucleic acid homologies among strains of Bacteroides melaninogenicus and related species

Saccharolytic, black-pigmented Bacteroides strains, which at present belong to the species Bacteroides melaninogenicus were classified on the basis of deoxyribonucleic acid (DNA) base ratios and DNA hybridization studies. These strains were divided into several DNA homology groups, which showed no or low mutual DNA homology. A DNA homology group with a percentage guanine plus cytosine (G + C) of 42–43% was formed by three strains of Bact. melaninogenicus subsp. melaninogenicus ; the type strain of this subspecies, strain ATCC 25845, had about 60% DNA homology with this group. Strain ATCC 15930, which has been assigned to this subspecies, had a percentage G + C of 47% and showed no DNA homology with the former group. All strains of Bact. melaninogenicus subsp. intermedius had a percentage G + C of 39–45%. A DNA homology group was formed by eight strains of this subspecies. The type strain of Bact. melaninogenicus subsp. intermedius , ATCC 25611, showed relatively low DNA homology with this main DNA homology group. A strain of Bact. melaninogenicus subsp. intermedius serotype C1 showed no DNA homology with the other strains tested. Furthermore two strains labelled 'Bact. melaninogenicus subsp. levii' were found to form a distinct DNA homology group. On the basis of the DNA homology results, the strains, which at present are classified in the species Bact. melaninogenicus , were clearly distinguished from strains of Bact. asaccharolyticus and Bact. gingivalis , and also from strains of related non-pigmented Bacteroides species.     

Survival of Escherichia coli with and without ColE1::Tn5 after aerosol dispersal in a laboratory and a farm environment.

The survival of a laboratory strain and a naturally occurring fecal strain of Escherichia coli, with and without a Tn5-containing derivative of ColE1, was examined after aerosol dispersal in a laboratory office and a barn under ambient temperature and humidity conditions. Following the release of paired strains, air and diverse types of surfaces were assayed for the test organisms. In both environments, the number of airborne bacteria declined rapidly within the first 2 h. Longer survival was found on surfaces and varied with surface type: recovery was greatest from wood products. Organisms persisted for 1 day in the office and for up to 20 days in the barn. Survival of the fecal strain was better than that of the laboratory strain in both test environments. In general, plasmid-bearing strains fared similarly to their plasmidless parents, but in several comparisons the ColE1::Tn5-containing strain showed enhanced survival. These studies have implications for the present and proposed release of genetically engineered organisms with and without plasmid vectors.     

Survival of Escherichia coli with and without ColE1::Tn5 after aerosol dispersal in a laboratory and a farm environment

The survival of a laboratory strain and a naturally occurring fecal strain of Escherichia coli, with and without a Tn5-containing derivative of ColE1, was examined after aerosol dispersal in a laboratory office and a barn under ambient temperature and humidity conditions. Following the release of paired strains, air and diverse types of surfaces were assayed for the test organisms. In both environments, the number of airborne bacteria declined rapidly within the first 2 h. Longer survival was found on surfaces and varied with surface type: recovery was greatest from wood products. Organisms persisted for 1 day in the office and for up to 20 days in the barn. Survival of the fecal strain was better than that of the laboratory strain in both test environments. In general, plasmid-bearing strains fared similarly to their plasmidless parents, but in several comparisons the ColE1::Tn5-containing strain showed enhanced survival. These studies have implications for the present and proposed release of genetically engineered organisms with and without plasmid vectors.     

Differing organic Acid exudation pattern explains calcifuge and acidifuge behaviour of plants

Many vascular plant species are unable to colonize calcareous sites. Thus, the floristic composition of adjacent limestone and acid silicate soils differs greatly. The inability of calcifuge plants to establish in limestone sites seems related to a low capacity of such plants to solubilize and absorb Fe or phosphate from these soils. Until now, mechanisms regulating this differing ability of plants to colonize limestone sites have not been elucidated. We propose that contrasting exudation of low-molecular organic acids is a major mechanism involved and show that germinating seeds and young seedlings of limestone plants exude considerably more di- and tricarboxylic acids than calcifuges, which mainly exude monocarboxylic acids. The tricarboxylic citric acid is a powerful extractor of Fe, and the dicarboxylic oxalic acid a very effective extractor of phosphate from limestone soils. Monocarboxylic acids are very weak in these respects. The study is based on ten species from limestone soils and ten species from acid silicate soils.     

Comparative analysis of extreme acid survival in Salmonella typhimurium, Shigella flexneri, and Escherichia coli.

Several members of the family Enterobacteriaceae were examined for differences in extreme acid survival strategies. A surprising degree of variety was found between three related genera. The minimum growth pH of Salmonella typhimurium was shown to be significantly lower (pH 4.0) than that of either Escherichia coli (pH 4.4) or Shigella flexneri (pH 4.8), yet E. coli and S. flexneri both survive exposure to lower pH levels (2 to 2.5) than S. typhimurium (pH 3.0) in complex medium. S. typhimurium and E. coli but not S. flexneri expressed low-pH-inducible log-phase and stationary-phase acid tolerance response (ATR) systems that function in minimal or complex medium to protect cells to pH 3.0. All of the organisms also expressed a pH-independent general stress resistance system that contributed to acid survival during stationary phase. E. coli and S. flexneri possessed several acid survival systems (termed acid resistance [AR]) that were not demonstrable in S. typhimurium. These additional AR systems protected cells to pH 2.5 and below but required supplementation of minimal medium for either induction or function. One acid-inducible AR system required oxidative growth in complex medium for expression but successfully protected cells to pH 2.5 in unsupplemented minimal medium, while two other AR systems important for fermentatively grown cells required the addition of either glutamate or arginine during pH 2.5 acid challenge. The arginine AR system was only observed in E. coli and required stationary-phase induction in acidified complex medium. The product of the adi locus, arginine decarboxylase, was responsible for arginine-based acid survival.     

Persistence of Escherichia coli O157 on farm surfaces under different environmental conditions

AIMS: To compare the persistence of Escherichia coli O157 on a variety of common faecally contaminated farmyard material surfaces (wood and steel) under different moisture and temperature regimes. METHODS AND RESULTS: Samples of field-conditioned farmyard materials (galvanized steel and wood) were cut into pieces and contaminated with fresh cattle faeces inoculated with nontoxigenic E. coli O157 (strain 3704). Thereafter, they were stored at four different environmental conditions; with temperature (5 and 20 degrees C) and moisture (moist or dry) as variables. Transfer of the pathogen to hands from the surfaces was also evaluated. Escherichia coli O157 numbers declined over time on all surfaces albeit at different rates according to the sample material and environmental conditions. Persistence was greatest on moist wood samples under cooler temperatures with large population numbers remaining after 28 days. Desiccation of surfaces resulted in a more rapid decline in E. coli O157 populations under both temperature regimes. Substantial numbers of colonies may also potentially be transferred to human hands from the surfaces during brief contact. CONCLUSIONS: When environmental conditions are favourable, E. coli O157 may persist for considerable times on a range of surfaces. However, when exposed to higher temperatures and dehydration, survival is notably decreased. Overall, bacterial persistence was significantly greater on wood samples relative to steel. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli O157 is a prevalent pathogen, common in ruminant faeces. Contact with contaminated faeces may lead to human infection, resulting in possible severe illness. Although our study used only one strain of bacteria, our findings indicates that E. coli O157 has the potential to persist for long periods of time on gates, stiles and other farmyard surfaces under a range of environmental conditions. These farmyard surfaces therefore pose a potential infection pathway particularly where there is a high risk of direct human contact (e.g. child petting zoos, open farms).     

Plant Seeds as Mineral Nutrient Resource for Seedlings--A Comparison of Plants from Calcareous and Silicate Soils

Mineral nutrients of seeds constitute a significant source ofessential elements to seedlings and developing individuals ofvascular plants. In spite of their potential ecological significance,seed nutrient pools have attracted little attention with respectto calcifuge–calcicole behaviour of plants. The objectivesof this study were, therefore, to compare concentrations of13 macro- and micronutrients (K, Rb, Mg, Ca, Mn, Fe, Co, Cu,Zn, Mo, B, P and S) in seeds and leaves of 35 mainly herbaceousvascular plant species growing on both limestone (calcareous)and silicate (non-calcareous) soils. Concentrations of Rb andCo in seeds of plants originating from limestone soils were,on average, about half of those from silicate soils. Concentrationsof Mn, Mg, Zn and P of seeds were, or tended to be, lower orslightly lower in limestone-soil plants, whereas mean Ca andMo concentrations were higher. Comparing seed and leaf concentrationsof the same species from limestone and silicate soils generallydemonstrated a high P enrichment ratio, but a particularly lowK enrichment ratio in seeds, valid for both types of soil. Itwas also apparent that Fe and Mn, micronutrients which are lessreadily solubilized and taken up by plants on limestone soils,had significantly higher seed:leaf concentration ratios in plantsfrom limestone than from silicate soils, whereas the oppositewas true for Ca. This indicates a ‘strategy’ tosatisfy the demand of seedlings for elements which are lessreadily available in the soil.Copyright 1998 Annals of BotanyCompany Seed, leaf, plant, nutrient, content, calcareous, silicate, acid, soil.